Abstract
The search for promising biomolecules such as chitooligosaccharides (COS) has increased due to the need for healing products that act efficiently, avoiding complications resulting from exacerbated inflammation. Therefore, this study aimed to produce COS in two stages of hydrolysis using chitosanases derived from Bacillus toyonensis. Additionally, this study aimed to structurally characterize the COS via mass spectrometry, to analyze their biocompatibility in acute toxicity models in vivo, to evaluate their healing action in a cell migration model in vitro, to analyze the anti-inflammatory activity in in vivo models of xylol-induced ear edema and zymosan-induced air pouch, and to assess the wound repair action in vivo. The structural characterization process pointed out the presence of hexamers. The in vitro and in vivo biocompatibility of COS was reaffirmed. The COS stimulated the fibroblast migration. In the in vivo inflammatory assays, COS showed an antiedematogenic response and significant reductions in leukocyte migration, cytokine release, and protein exudate. The COS healing effect in vivo was confirmed by the significant wound reduction after seven days of the experiment. These results indicated that the presence of hexamers influences the COS biological properties, which have potential uses in the pharmaceutical field due to their healing and anti-inflammatory action.
Highlights
Chitin, in addition to being a valuable component in arthropod exoskeletons, is present in fungal cell walls and can be used as a raw material to obtain other substances [1].For this, it is necessary to convert this molecule into certain derivatives, which are of great scientific interest in relation to their effects in several fields of research [2]
As the anti-inflammatory and healing actions of COS produced through chitosanases obtained from Bacillus toyonensis have still not been reported in the literature, the results found here will contribute to demonstrating the potential of these molecules, highlighting the importance of using residues rich in chitin from crustaceans as sources of bioactive components in the pharmaceutical field
The Bradford method was used to quantify the protein plasma extravasation induced by the zymosan administration (s.c.) in the air pouch formed on the animal’s back; the results demonstrated that all doses of COS1 and COS5 significantly reduced the protein concentration in the air pouch when compared to the group that received only saline solution (i.g.) and zymosan (s.c.) (Figure 8A)
Summary
In addition to being a valuable component in arthropod exoskeletons, is present in fungal cell walls and can be used as a raw material to obtain other substances [1].For this, it is necessary to convert this molecule into certain derivatives, which are of great scientific interest in relation to their effects in several fields of research [2]. Studies indicate that COS have multiple biological properties, such as anti-inflammatory, immunostimulant, antimicrobial, antitumor, antidiabetic, antioxidant, and neuroprotective effects [15,16,17,18,19,20,21,22,23]. With regard to their healing activity, different in vitro and in vivo studies demonstrate the potential of COS; most of these reports consist of the evaluation of these substances together with other products and few have described their action in animal healing trials [24,25]. Further studies are needed to better elucidate their effects when repairing skin wounds
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